PROGRAM SUMMARY The current Official US Standards for Grains regulatory guidelines for ergot contamination in grains is a crude measurement and determines the percentage of ergot sclerotia in a sample, with ergoty grain defined as > 0.05% sclerotia in wheat, > 0.1% sclerotia in barley, oats, and triticale, and > 0.3% sclerotia in rye (CFR Title 7). The European Union Scientific Panel on Contaminants in the Food Chain concluded that a consistent relationship between the alkaloid concentration in the sclerotia and total ergot alkaloid concentration could not be established (EFSA 2005). No country has limits for individual ergot alkaloids in food or feed. Several recent factors have increased ergot invasion in crops, including no-till farming practices leaving the vegetation and ergot sclerotia on the soil surface for increased infection rates, the move to organic farming and avoiding fungicides, and the dramatic weather patterns with huge rainfalls that enhance increased Claviceps purpurea invasion of cereals and grasses in the spring/summer. No US veterinary diagnostic laboratory is providing a validated ergot alkaloid analysis of cereal grains and grasses by LC/MS/MS for the main Claviceps ergot alkaloids, the ergopeptines (- ines, biologically active) include ergotamine, ergosine, ergocristine, ergocryptine, and ergocornine, and their diastereomers the ergopeptinines (-inines, biologically inactive). In nature, ergopeptinines always accompany ergopeptines, and during storage of raw materials over time or improper storage, more ergopeptinines may form. The conversion of -ines to -inines can be rapid under certain laboratory conditions and -inines can be reactivated to -ines under certain conditions. Therefore, both epimers have to be determined for ergot alkaloid contamination of foods and feeds (Krska and Crews). This proposal is to develop and validate a sensitive LC/MS/MS method for ergpopeptines, ergotamine, ergosine, ergocristine, ergocryptine and ergocornine, and their -inines in cereal grains of wheat, barley, rye and grasses. The inclusion of a grass matrix for ergot alkaloid analysis by LC/MS/MS is not commonly addressed in any method but is extremely important for veterinary livestock exposures. The analytical range of detection chosen for the assay, 20 to 8000 ug/kg, represents veterinary toxicological important data for interpretation of animal feed and food contamination. RELEVANCE The LC/MS/MS analytical method for ergopeptines and ergopeptinines in small cereals and grasses will provide chemical analysis of potentially contaminated feed to be used for food animals or sold into the food chain. The current US Official Grain Standards of determining percentage contamination by ergot sclerotia in cereal grains is often inaccurate and ignores the great variation of alkaloid content in individual sclerotia. The increased use of no-till farming and leaving vegetation and sclerotia on the soil surface for the successive crops, consumer desire to utilize organic foods without chemicals, and dramatic weather events will tend to increase the incidence of Claviceps purpurea in cereal crops and grasses and exposure to ergot alkaloids.